GSM7384139: Ag challenge - BT1; Pseudomonas aeruginosa; RNA-Seq
1ml of samples were centrifuged at 3,500 X g for 10 min to pellet the cells. The cell density here was 1.5 X 108CFU/ml bacteria. Zirconia beads were used for cell lysis for at least 2 min using a bead beater instrument. The total RNA was then extracted with a RiboPure Bacteria RNA Isolation Kit (Invitrogen, USA) according to the manufacturer's recommendations. To ensure that the DNA was completely removed, residual DNA was removed with DNase (Sigma-Aldrich Inc, St Louis, MO, USA) to get a pure RNA sample. Libraries were prepared using the New England Biolabs NEBNext rRNA depletion bacteria kit and NEBNext Ultra II Directional RNA library prep kit for Illumina, according to the manufacturer's instructions.